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Biomedical subjects

K Takamori

Publications and source records attributed to K Takamori.

At least 19 recordsLinked to original sources

Organ culture conditions of human hair follicles.

Experimental results revealed that although [3H]thymidine uptake in the hair bulb increased time dependently for 12 days under normal culture conditions (95% air-5% CO2 at 37 degrees C), striking morphological changes occurred in the hair bulb cells as demonstrated by histological findings. As such, organ culture conditions applicable to human hair follicles were studied utilizing observations from both histology and DNA synthesis. We found that culture conditions of 95% O2-5% CO2 at 31 degrees C were superior when compared to normal culture conditions for cultures of human hair follicles when attempting to maintain the normal morphology of hair germinative cells. The hair bulb and the germinative cells successfully maintained their normal morphology throughout the 96 and 48 h culture period, respectively. Autoradiographs of [3H]thymidine-labeled follicles showed localization in the germinative cells below Auber's critical line. Hair bulb DNA synthesis increased time dependently for 96 h after culture initiation. Under conditions of 95% O2-5% CO2 at 31 degrees C, the synthesis of DNA in hair germinative cells was observed. Such an organ culture method may prove useful for studies on the human hair growth mechanism.

Autoradiography

Increased HLA-DR+ T-lymphocyte population in peripheral blood of alopecia areata.

The populations of activated T-cell subsets [HLA-DR(+)-Leu 4+ cells, interleukin 2 receptor positive (IL-2R+)-Leu 4+ cells] in the peripheral blood of patients with alopecia areata (AA) were investigated using double direct immunofluorescence staining. Fifty-eight patients with AA were classified into one of three types: those with inactive single AA (type 1) lesions, active multiple alopecia areata (MAA) lesions and active alopecia totalis (AT) (type 2) and chronic alopecia universalis (AU) (type 3). Compared to normal controls, high percentages of HLA-DR(+)-Leu4+ cells were detected in types 2 and 3 AA patients, but not in type 1 AA patients. These findings suggest that T cells are activated in the peripheral blood of active MAA, AT and chronic AU.

Adult

[Three-dimensional MR brain image using volume rendering method].

3-dimensional image reconstructed from the data of computer tomography has provided useful informations for the pre-operative evaluation in the field of neurosurgery and plastic surgery. However CT has failed to visualized the 3-dimensional surface image of the central nervous system. On the other hand the progress of high-speed data acquisition technique and 3-dimensional fourier transformation has enabled the practical 3-dimensional image using magnetic resonance imager. However for the visualization of a pure surface image of the brain and spinal cord a sophisticated method and special image work station has been necessary. The authors describe a technique for the pure surface image of the brain and spinal cord using a normal mini-computer system and magnetic resonance imager. This image has a good correlation to the macroscopic anatomical brain surface (sulci and gyri).

Brain

[The partial characterization of neutrophils' and lymphocytes' chemotactic factors from murine anagenic hair bulb extract].

Sephadex G-75 gel filtration of murine anagenic hair bulb extracts (HBE) showed two peaks with chemotactic activity. Peak I (m.w. 63 +/- 4.6 KD) was chemotactic for both neutrophils and lymphocytes, and Peak II (m.w. 47 +/- 5.6 KD) was chemotactic for lymphocytes only. The neutrophil related chemotactic activity was sensitive to treatments by trypsin, pronase, neuraminidase, or heating at 100 degrees C for 10 min. With Peak I, lymphocyte related chemotactic activity was sensitive to treatments by pronase, trypsin or neuraminidase, but was unaffected by heating at 100 degrees C for 10 min. On the other hand, the same aforementioned treatments all individually inactivated the Peak II lymphocyte related chemotactic activity. Intradermal injection of Peak I fractions into guinea pigs induced infiltration of neutrophils and mononuclear cells, and that of Peak II induced the infiltration of mononuclear cells. These findings suggest that normal C3H murine anagenic hair bulb contains three different chemotactic factors related to neutrophils and lymphocytes.

Animals

Purification and characterization of an extracellular proteinase from Hendersonula toruloidea.

An extracellular proteinase from a fast-growing strain of Hendersonula toruloidea was demonstrated when it was cultivated in liquid medium containing bovine serum albumin as the sole nitrogen source. Purification to homogeneity of the proteinase was performed by carboxymethyl cellulose, CM Sephadex G-50, and Sephacryl S-200 column chromatographies. The purified enzyme was a chymotrypsinlike serine proteinase, as indicated by the strong inhibitory activities of diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, L-1-tosylamido-2-phenylethylchloromethyl ketone, and chymostatin and good kinetic constants for a synthetic substrate, Suc-Ala-Ala-Pro-Phe-MCA. The enzyme had a pI of 8.4, a pH optimum of 9.0, and a molecular weight of 34,000. Skin constituents such as stratum corneum and nail, but not hair, were easily digested by this enzyme. Thus, this extracellular proteinase may play a role in the invasion of thickly keratinized skin and nail by this organism.

Amino Acid Sequence

[A Japanese family of Nettleship Falls X-linked ocular albinism].

A family with Nettleship Falls X-linked ocular albinism (NFXOA) was reported. The funduscopic examination revealed that both the male proband case and his maternal male cousin with nystagmus and amblyopia had macular hypoplasia and slightly less pigmented fundus than normal Japanese, and the carrier female (proband's mother) had the characteristic pigmentary mosaicism of the fundus. The slitlamp biomicroscopic examination revealed that the proband case's iris was less pigmented and the carrier female had the normal coloured iris. Translucency was not found in the iris of either the proband case or the carrier female. Hypopigmented maculas were found on the skin of the proband case. Histopathological findings of the cutaneous tissue of both the proband case and the carrier female showed macromelanosomes in the epidermis. The diagnosis of NFXOA is considered to be difficult by ocular clinical signs in the case of Japanese male patients without both characteristic albinotic fundus and the mother's pigmentary mosaicism of the fundus. Macromelanosomes recognized by skin biopsy are helpful for diagnosis.

Adult

[The purification of high molecular weight protease from the rat's skin].

In this study, the purification of a high molecular weight protease from Wistar Rats' skin was investigated. A high molecular weight protease from Wistar Rats' skin was purified by successive (NH4)2SO4 fractionation, Phenylsepharose CL-4B, and HPLC gel filtration chromatographies. The activity of the enzyme was determined using a synthetic substrate, succinyl-leucyl-leucyl-valyl-tyrosine methylcoumarinamide (SLLVT-MCA). The molecular weight of the purified enzyme was estimated to be 750,000 by gel filtration. With SDS-PAGE the purified enzyme was dissociated into several bands of which the majority fell into the range of 20,100-36,000 daltons. These findings suggest that high molecular weight protease in rat's skin consists of various subunits.

Animals

[A study on the therapy and prognosis of pemphigus vulgaris].

The treatment and, in particular, the prognosis of 32 patients with pemphigus were studied. All cases were classified according to treatment, for example, corticosteroid only, corticosteroid + immune-suppressive agents, plasma exchange + corticosteroid and/or immune-suppressive agents, and other treatments. Results showed that with the plasma exchange group skin lesions disappeared the quickest, the percentage of disappearance was the highest and a long standing remission was able to be maintained. With the plasma exchange group, many cases were severe and had complications which were not able to be resolved by corticosteroid or immune-suppressive agents. Therefore, plasma exchange may be considered as a very significant form of treatment of pemphigus vulgaris.

Adrenal Cortex Hormones

[The partial characterization of neutrophils and lymphocytes chemotactic factors from murine anagenic hair bulb extract].

The murine anagenic ++ hair bulb extract (HBE) showed a chemotactic activity for both neutrophils and lymphocytes in vitro. The activity for both cells was retained after heating at 56 degrees C for 30 min, and dialysis. However, heating at 100 degrees C for 10 min inactivated the chemotactic activity for neutrophils, but not for lymphocytes. Activity for neutrophils was sensitive to degradation by trypsin or pronase but stable to neuraminidase treatment. On the other hand, activity for lymphocytes was not affected by protease treatment, but was reduced by approximately 30% after being treated by neuraminidase. These findings suggest that HBE contains at least two kinds of chemoattractants for neutrophils and lymphocytes.

Animals

[Skin high-molecular-weight protease. 2. Biochemical properties].

The purification of High-Molecular-Weight Protease from Wistar Rat's skin was previously investigated and reported. Some properties of this enzyme were subsequently investigated. This enzyme showed high activities toward succynyl-leucyl-leucyl-valyl-tyrosine-methylcoumarinamide (SLLVT-MCA). The addition of 0.03% sodium dodecyl sulphate (SDS) stimulated enzyme activity to 2,000% of the initial activity. The pH optimum was pH 8.5 in the presence of 0.03% SDS, but pH 6.0-8.5 in the absence of SDS. This enzyme was inhibited by one of serine protease inhibitors, diisopropylfluorophosphate (DFP) strongly. It was also inhibited by some of SH protease inhibitors, N-ethylmaleimide (NEM), iodoacetamide (IA), and leupeptin. Interestingly, chymostatin, a serine and SH protease inhibitor also inhibited enzyme activity. These results show that both serine and cysteine++ residues are related to the enzyme activity of High-Molecular-Weight Protease.

Animals

Differences in induction of lysosomal protease activity by protease inhibitors in B16 melanoma cell lines.

1. The effects of potent protease inhibitors in vitro (leupeptin, pepstatin and E-64[N-[L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]agmatine]) on intracellular cathepsin B (EC 3.4.22.1), hemoglobin (Hb)-hydrolase and acid phosphatase (EC 3.1.3.2) from cultured B16 melanoma variants (B16-F1, F10 and BL6) were studied. 2. E-64 induced all the cultured B16 melanoma variants to decrease the activity of intracellular cathepsin B but did not have this effect with Hb-hydrolase or acid phosphatase. Furthermore, E-64 decreased the activity of cathepsin B in both the lysosomal and cytosol fractions. 3. Leupeptin induced all the cultured B16 melanoma variants to increase the activities of intracellular cathepsin B and Hb-hydrolase but not that of acid phosphatase. An increase in the level of cathepsin B activity was most significant in B16-BL6 followed by F10 and then F1 variants. 4. Leupeptin induced all the cultured B16 melanoma variants to increase the cathepsin B activity in the lysosomal fraction. Our data differed from the results of Tanaka et al. (1981) in that leupeptin induced rat cultured hepatocytes to inhibit the activity of intracellular cathepsin B and increase the Hb-hydrolase activity, especially in the cytosol fraction.

Acid Phosphatase

Interaction of tumor and surrounding tissue of mice inoculated B16 melanoma variants in terms of enzyme activity.

1. The interactions of B16-F1 and B16-F10 tumors with their surrounding tissues in terms of enzyme activities such as cathepsin B, hemoglobin(Hb)-hydrolase, acid phosphatase, beta-glucuronidase and plasminogen activator were investigated when said tumors proliferated locally and at secondary sites throughout the host's circulatory system. 2. In the case of B16-F1 and B16-F10 tumor cells proliferating under the skin, statistical differences were not detected between the enzyme activities of the skin surrounding the tumors and control skin, nor between B16-F1 and B16-F10 tumors, except for beta-glucuronidase. 3. In the case of B16-F1 and B16-F10 tumor cells metastasizing to lung, statistical differences were detected between numerous enzyme activities of the lung tissues surrounding the tumors and control lung tissue, and also between B16-F1 and B16-F10 tumors. 4. The activities of cathepsin B and acid phosphatase of lung tissue surrounding B16-F1 tumor were lower than those of the control lung. 5. beta-Glucuronidase activity of lung tissue surrounding B16-F10 tumor was higher than that of the control lung. 6. The activities of cathepsin B, Hb-hydrolase and beta-glucuronidase of the B16-F10 tumor were higher than those of the B16-F1 tumor. 7. Results indicate that metastasized B16 melanoma tumor cells interact with surrounding lung tissues, and that cathepsin B, Hb-hydrolase and beta-glucuronidase might play important roles in the metastasis of the malignant tumor.

Acid Phosphatase

Isolation of a keratinolytic proteinase from Trichophyton mentagrophytes with enzymatic activity at acidic pH.

A keratinolytic proteinase with enzyme activity at acidic pH was isolated from culture filtrates of Trichophyton mentagrophytes, a major pathogenic fungus of dermatophytosis. The molecular weight of the proteinase was estimated to be 41,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 38,000 by gel filtration. The isoelectric point was determined to be 3.9. The proteinase had a pH optimum of 4.5 for keratin and 5.5 for hemoglobin. This enzyme hydrolyzed the synthetic chymotrypsin substrate Suc-Ala-Ala-Pro-Phe-MCA (Km, 0.59 mM), and its activity was strongly inhibited by chymostatin. Previously reported proteinases from dermatophytes have had enzyme activities in neutral or alkaline pH; however, healthy skin has a weakly acidic pH. Thus, the purified proteinase which has an optimal activity at acidic pH and hydrolyzes skin constituents could be an important virulence factor in dermatophytosis.

Cations, Divalent

[Increase in Leu7- Leu11+ cells in the peripheral blood lymphocytes of patients with alopecia areata].

Natural killer (NK) cell subsets in the peripheral blood lymphocytes of patients with alopecia areata (AA) were investigated by FACStar-I using NK cell specific monoclonal antibodies. The patients were classified into one of 2 types (Type A: single AA in regrowth phase; Type B: multiple AA, alopecia totalis and alopecia universalis in non-regrowth phase). The percentages of Leu7- Leu11+ cells in Type B were significantly higher than those of the normal control. The increase in Leu7- Leu11+ cells in Type B suggests that NK cells may play an important role in the pathogenesis of AA in the non-regrowth phase.

Adult

[Correlation of activated T cells and natural killer (NK) cells with the disease activity of alopecia areata (AA)].

Patients with AA were classified into one of two types (Type A: active AA with less than 3 patches; Type B: active AA with 10 or more patches). The percentages of activated T cell subsets and NK cell subsets in the peripheral blood of Type A were similar to those of the normal control. On the other hand, the percentages of HLA-DR+ Leu2a+ cells, HLA-DR+ Leu3a+ cells, Leu7+ Leu11+ cells and Leu7- Leu11+ cells in Type B were significantly higher than those of the normal control. These findings suggest that activated T cells and NK cells play an important role in the disease activity of AA.

Adult

[Reaction mechanism catalyzed by skin sulfhydryl oxidase].

The mechanism involved in the reaction catalyzed by skin sulfhydryl oxidase was investigated. Skin sulfhydryl oxidase, which was purified to a 300-fold increase in specific activity over rat skin, was used. As the substrate for the reaction catalyzed by skin sulfhydryl oxidase, dithiothreitol (DTT) was used. The disappearance of DTT, consumption of O2 and production of H2O2 were measured according to Takamori's Janolino's and Kakinuma's methods, respectively. The molar ratio of the disappearance of DTT and the consumption of O2 during the enzyme reaction was 1:1.05. The molar ratio of the disappearance of DTT and the H2O2 production during the enzyme reaction was 1:086. Results therefore showed that the molar ratio of the disappearance of DTT, consumption of O2, and production of H2O2 was 1:1:1. Therefore, the stoichiometry of the enzyme reaction was estimated to be (formula; see text)

Animals

Partial purification and characterization of alpha-glucosidase from Rothia dentocariosa.

Disaccharidases of oral bacteria, especially alpha-glucosidase and beta-fructofuranosidase, are considered to play an important role in the induction of dental caries. Upon the examination of disaccharidases from several strains of saccharolytic oral bacteria, we found all of those bacteria to be capable of hydrolyzing the glycosidic linkage of sucrose. One species of bacteria, Rothia dentocariosa, was found to contain a single disaccharidase, alpha-glucosidase. This enzyme was partially purified by ammonium sulfate precipitation, gel filtration and ion-exchange column chromatography. The optimum pH and temperature for the enzyme activity was found to be 6.8-7.0 and 40 degrees C, respectively. The enzyme activity was strongly inhibited by Ag+, Hg2+, Cu2+, Fe2+ and Tris (Hydroxymethyl) aminomethane.

Bacteria